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doi:10.1534/genetics.105.042333
A more recent version of this article appeared on October 1, 2005.
REGULAR RESEARCH PAPERS |
Mcm10 is Required for the Maintenance of Transcriptional Silencing in Saccharomyces cerevisiae
Ivan Liachko 1 and Bik K Tye 1*
1 Cornell University
* To whom correspondence should be addressed. E-mail: bt16{at}cornell.edu.
Submitted on February 21, 2005
Revised on March 29, 2005
Accepted on 27 June 2005
Mcm10 is an essential protein that participates in both the initiation and elongation of DNA replication. In this study we demonstrate a role for Mcm10 in the maintenance of heterochromatic silencing at telomeres and HM loci of budding yeast. Two mcm10 mutants drastically reduce silencing of both URA3 and ADE2 reporter genes integrated into these silent loci. When exposed to alpha-factor, mcm10 mutant cells display a "shmoo-cluster" phenotype associated with a defect in the maintenance of silencing. In addition, when combined with a defect in the establishment of silent chromatin, mcm10 mutants demonstrate a synergistic defect in HML silencing. Consistent with a direct silencing function, Mcm10p shows a two-hybrid interaction with Sir2p and Sir3p that is destroyed by the mcm10-1 mutation and dependent on the C-terminal 108 amino acids. Tethering GBD-MCM10 to a defective HMR-E silencer is not sufficient to restore silencing. Furthermore, mutations in MCM10 inhibit the ability of GBD-SIR3 to restore silencing when tethered to a defective HMR-E. Suppressor mutations in MCM2, which suppress the temperature sensitivity of mcm10-1, fail to overcome the mcm10-1 silencing defect suggesting that MCM10's role in transcriptional silencing may be separate from its essential functions in DNA replication.
Key Words: DNA replication factor Mcm10, heterochromatin maintenance, silencing of HML and HMR, silent information regulator proteins Sir2 and Sir3, telomeric silencing
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